These include 9 techniques
(1) Standard polymerase chain reaction (PCR):
Given Nobel Prize in Chemistry in 1993.
@ He was able to amplify DNA by repeated
cycles driven by DNA polymerase enzyme.
@ PCR technique used an automated process
for DNA amplification & separation of the
DNA strands.
PCR Applications:
1. DNA extraction: From cell genome DNA by
selective amplification of a specific region
of DNA. This is done by hybridization ((تهجين
probe and DNA clones (نسيلات) techniques to
create copies of DNA fragments.
2. DNA sequencing: To determine unknown
PCR-amplified sequences by the insertion of
a DNA sequence into a genetic material of
another organism.
3. DNA-based phylogeny: To determine the
evolutionary relationships among organisms.
Extracted DNA
4. Identification of uncultural or slow-growing
organisms:
Such as mycobacteria,
anaerobic bacteria, or viruses.
5. Early diagnosis of infectious diseases:
Such as leukemia and lymphomas; detecting
malignant cells at a high sensitivity.
PCR
6. Quantification viral load in a patient (HIV)
7. Diagnosis of hereditary diseases
8. Identification of genetic fingerprints. This
is used in forensic sciences and paternity
testing
Paternity fingerprints (1) Father. (2) Child. (3) Mother.
(2) Real-time PCR:
@ Real time means immediate. Also called
quantitative PCR (qPCR)
@ It is used to amplify and quantify DNA.
@ qPCR enables at the same time both
detection and quantification.
@ qPCR technique uses hybridization probe
technique.
DNA probe hybridization
Applications of qPCR:
1.DNA amplification, DNA quantification,genotyping.
2. Quantification of microbial load in foods and
vegetables.
3. Rapid detection of cancer, newly emerging
diseases, and genetic abnormalities.
4. Microbial assessment of water quality.
5. Production of plant seedlings that are free
of pathogens.
PCR cycler
(3) Dna Sequencing & Genome Data Technique:
Used to analyze the bacterial DNA sequence
and mapping all its genome data.
(4) Rna Interference Technique:
Used in biological research and drug discovery
(5) 16s Rrna Sequencing Technique:
Used for bacterial identifications
Automated Genetic Analyzer for completegenome sequencing .
(6) Gel Electrophoresis Technique:
Used to separate DNA, RNA, or protein
molecules by blotting through an electric
field by virtue of their size, shape or
electric charge.
Gel electrophoresis machine
(7) Dna Microarray Technique:
@ Used as an alternative to the electrophoresis
blotting techniques.
@ This technique can detect the presence of
pathogens in a sample.
@ It can determine the genetic differences
between two microbial strains.
Microarrays Machine
(8) Direct Gene Detection Technique:
Used to detect genes related to drug
resistance mechanisms, such as mecA gene
in Staphylococcus aureus
Gene detection
(9) Pulsed-field gel electrophoresis Technique:
@ Used for separation of large DNA molecules
by applying to a gel matrix electric field that
periodically changes direction.
@ It is similar to standard gel electrophoresis
except that instead of running the current
in one direction, it is run in 3 directions.
@ Used for genotyping, genetic fingerprinting,
and tracing sources of infections.
Genotyping technique
